Microscopic and molecular detection of piroplasms among sheep in Upper Egypt.

Introduction: Blood parasites pose a significant threat to livestock production in southern Egypt, yet there is a scarcity of information regarding their circulation and epidemiology in sheep in this region. This study aimed to investigate the seroprevalence of blood parasite infections in sheep in Assiut governorate, Upper Egypt. Methods: A total of 400 blood samples were collected from sheep of varying ages and genders. The preliminary screening for the presence of piroplasms, mainly Babesia and Theileria spp., via microscopic examination, followed by investigation of the potential risk factors linked with the exposure to infection. Moreover, molecular identification of both parasites on some of positive samples was performed using PCR targeting Babesia 18S rRNA and Theileria annulata Tams1 gene. Results: The microscopic examination revealed that among the examined sheep, there was an overall prevalence of blood parasites at 44% (176 out of 400), with Babesia spp. observed in 14% (56 out of 400) and Theileria spp. in 30% (120 out of 400). Furthermore, the infection rate was non-significantly higher in young animals (50%) compared to adults (38.5%) (P = 0.246). Male sheep exhibited a significantly higher vulnerability to both parasites' infection (63.3%) compared to females (35.7%) (P = 0.011). Interestingly, the prevalence of both blood parasites was significantly higher during the cold season (66.1%) compared to the hot season (15.9%) (P = < 0.001). The molecular analysis identified the presence of Babesia ovis and Theileria annulata among a subsample of the positive sheep's bloods films. The identified species were recorded in the GenBank™ databases and assigned specific accession numbers (OQ360720 and OQ360719 for B. ovis), and (OP991838 for T. annulata). Conclusions: Taken together, this study confirms a high prevalence of piroplasmosis and offers epidemiological and molecular insights into blood parasites in sheep from Upper Egypt, highlighting the importance of detecting these parasites in various hosts and their competent vectors (ticks).

Molecular identification of Haemonchus contortus in sheep from Upper Egypt.

Introduction: Haemonchus spp. are considered the most important strongylid nematodes with a worldwide distribution. The parasite's blood-sucking nature can lead to severe anemia in infected animals. Despite its widespread impact, there is a dearth of comprehensive data on morphological and molecular identification methods for Haemonchus spp. in sheep from Upper Egypt. To address this gap, our current study aimed to assess the prevalence of Haemonchus spp. in 400 sheep fecal samples. Methods: We employed microscopic examination and molecular techniques, using polymerase chain reaction (PCR) targeting the 18S gene for precise identification. Additionally, the potential risk factors associated with the infection by the parasite in sheep were explored. Results: The study pointed out that 33.00% (132 of 400) of the examined sheep were infected with Haemonchus spp. Sheep age and seasonal variability were found to be significant factors (p < 0.05) associated with the infection. Notably, sheep under 2 years old exhibited a higher risk, with an infection rate of 43.75% (84 out of 192), than their older counterparts. Furthermore, all reported infections were exclusively observed during the cold season, constituting 58.93% (132 out of 224) of cases. By contrast, no statistically significant association (p > 0.05) was found between the sex of the examined sheep and the occurrence of haemonchosis. Employing molecular methods, we isolated and identified the parasite through PCR analysis of cultured larvae, which were then subsequently confirmed as Haemonchus contortus via phylogenetic analysis. Discussion: The study concluded that there was a relatively high occurrence of H. contortus among sheep from Upper Egypt. We recommend the implementation of stringent and effective control measures to combat the infection and safeguard livestock health.

The antimicrobial, antibiofilm, and wound healing properties of ethyl acetate crude extract of an endophytic fungus Paecilomyces sp. (AUMC 15510) in earthworm model.

The endophytic fungus Paecilomyces sp. (AUMC 15510) was isolated from healthy stem samples of the Egyptian medicinal plant Cornulaca monacantha. We used GC-MS and HPLC analysis to identify the bioactive constituents of ethyl acetate crude extract of Paecilomyces sp. (PsEAE). Six human microbial pathogens have been selected to evaluate the antimicrobial activity of PsEAE. Our data showed that the extract has significant antimicrobial activity against all tested pathogens. However, the best inhibitory effect was observed against Bacillus subtilis ATCC 6633 and Pseudomonas aeruginosa ATCC 90274 with a minimum inhibitory concentration (MIC) of 3.9 µg/ml and minimum bactericidal concentration (MBC) of 15.6 µg/ml, for both pathogens. Also, PsEAE exerts a significant inhibition on the biofilm formation of the previously mentioned pathogenic strains. In addition, we evaluated the wound healing efficiency of PsEAE on earthworms (Lumbricus castaneus) as a feasible and plausible model that mimics human skin. Interestingly, PsEAE exhibited a promising wound healing activity and enhanced wound closure. In conclusion, Paecilomyces sp. (AUMC 15510) could be a sustainable source of antimicrobial agents and a potential therapeutic target for wound management.

A comparative study of the toxic effect of ZIF-8 and ZIF-L on the colonization and decomposition of shaded outdoor mice carrions by arthropods.

Metal-organic frameworks (MOFs) are promising materials for several applications. Thus, they have been intensively reported and commercialized by several international companies. However, little is known about the fate and risk of MOFs to living organisms. Here, the toxic effect of two Zinc (Zn)-based MOFs; zeolitic imidazolate frameworks (ZIF-8) and leaf-like ZIF (ZIF-L), was tested to investigate the impact of the postmortem period of mice carrions and arthropods which found in decomposing carrions. The data analysis revealed an increase in zinc content over time. Toxicology in forensics studies biological materials for the presence of poisons, such as pharmaceuticals. The toxicology report can provide important details about the types of chemicals present in a person and whether the amount of those substances is in line with a therapeutic dose or exceeds a dangerous level. These findings conclude the possible fate and impact after mortality. This study presents the first study of the toxic effect of ZIFs materials using mice carrions and arthropods (Sarcophaga sp. Larvae) via morphological and microscopic studies compared with control, providing important biological information could aid in the environmental impact of the toxic level of MOF materials.

Antibacterial and wound-healing potential of PLGA/spidroin nanoparticles: a study on earthworms as a human skin model.

Aim: The essential protein element of spider silk 'spidroin' was used to assess its impact on the wound-healing process. Methods: Spidroin nanoparticles (NPs) were prepared using poly(lactic-co-glycolic acid) polymer (PLGA/spidroin NPs) at different weight ratios (5:1, 10:1, 15:1) and were in vitro characterized. The optimized NPs were tested in vitro for release and antibacterial activity. To assess wound-healing effects, NPs were topically applied on surgically induced injuries in Allolobophora caliginosa earthworms as a robust human skin model. Results: Optimized NPs (173 ± 3 nm) revealed considerable antibacterial effect against Staphylococcus aureus and Escherichia coli. After 4 days of NPs application on wounds, macroscopical and histological examinations revealed a significant reduction in wound and re-epithelialization times. Conclusion: PLGA/spidroin NPs may represent a promising option for wound repair.

The assessment of the protective impact of spidroin extract against UV-A radiation damage by using earthworms (Aporrectodea caliginosa) as a robust human skin model via macroscopic and histological observations.

Numerous studies have confirmed the damage caused by excessive exposure to ultraviolet-A rays. Malignant melanoma and skin cancer are two of the most serious health consequences. Thus, the UV-A protectant is intended to protect the skin, especially the two primary layers of skin (epidermis that represents the interface between the body and its surroundings and dermis). Spider silk is the most powerful natural fibre due to its regeneration, biocompatibility, antimicrobial, wound healing, antiseptic, and blood clotting properties. This work targeted to determine the protective effect of spidroin extract against UV-A radiation damage. Earthworms Aporrectodea caliginosa were collected from Assiut University's farm. Each set of ten earthworms was separated into six groups and placed in a plastic container. Webs of spiders collected from trees and old houses. Spidroin was extracted and utilised in this work to determine the potential effects of topical application on UV-A protection. The experiment is divided into two sections: (1) UV-A exposure and (2) the use of spidroin extract to protect the earthworms from ultraviolet radiation. Two control groups (1،2) of worms were not received UV-A exposure, and four groups (3,4,5,6) were exposed to UVR-A. In contrast, groups (5,6) were received spidroin extract before exposure to UV-A. Each group from the groups (3,4,5,6) was exposed for three consecutive days (» hour/day, ½ hour/day, and 1 h/day), using a UV-Lamp with a wavelength of 366 nm. The histopathological changes revealed that after 1/4 h of UV exposure, the cuticle was swollen with a slightly detached epithelium. The cuticle was down after 1/2 h of exposure, and the epidermis was totally damaged and necrosed. After 1 h, the exposure showed destruction of the epidermis in the circular muscle with a loss of muscle filament integrity, varying size, and altered nucleus form, along with mild disintegration of longitudinal muscle. Spidroin extract is critical for earthworm protection against UV-A radiation damage and able to regeneration. For the first time, morphological and histological analysis was established to detect the Spidroin extract evaluated for topical application on earthworms. Earthworms can be considered as a robust human skin model prior to UV-A exposure. It induces a complete protective effect against UV-A radiation damage in earthworms.

Coexisting mechanisms of bacterial community are changeable even under similar stable conditions in a chemostat culture.

The coexisting mechanism of a synthetic bacterial community (SBC) was investigated to better understand how to manage microbial communities. The SBC was constructed with three kinds of phenol-utilizing bacteria, Pseudomonas sp. LAB-08, Comamonas testosteroni R2, and Cupriavidus sp. P-10, under chemostat conditions supplied with phenol as a sole carbon and energy source. Population densities of all strains were monitored by real-time quantitative PCR (qPCR) targeting the gene encoding the large subunit of phenol hydroxylase. Although the supply of phenol was stopped to allow perturbation in the SBC, all of the strains coexisted and the degradation of phenol was maintained for more than 800 days. The qPCR analyses showed that strains LAB-08 and R2 became dominant simultaneously, whereas strain P-10 was a minor population. This phenomenon was observed before and after the phenol-supply stoppage. The kinetic parameters for phenol of the SBC changed before and after the phenol-supply stoppage, which suggests a change in functional roles of strains in the SBC. Transcriptional levels of phenol hydroxylase and catechol dioxygenases of three strains were monitored by reverse-transcription qPCR (RT-qPCR). The RT-qPCR analyses revealed that all strains shared phenol and survived independently before the phenol-supply stoppage. After the stoppage, strain P-10 would incur the cost for degradation of phenol and catechol, whereas strains LAB-08 and R2 seemed to be cheaters using metabolites, indicating the development of the metabolic network. These results indicated that it is important for the management and redesign of microbial communities to understand the metabolism of bacterial communities.

Draft Genome Sequence of Phenol-Degrading Variovorax boronicumulans Strain c24.

We report the draft genome sequence of Variovorax boronicumulans strain c24, which was isolated from a soil-inoculated chemostat culture amended with phenol as a sole carbon and energy source. The genome data will provide insights into phenol and other xenobiotic compound degradation mechanisms for bioremediation applications.

Draft Genome Sequence of Phenol-Degrading Variovorax boronicumulans Strain HAB-30.

We report the draft genome sequence of Variovorax boronicumulans strain HAB-30, which was isolated from a phenol-degrading chemostat culture. This strain contains genes encoding a multicomponent type of phenol hydroxylase, with degradation pathways for catechol and other aromatic compounds. The genome sequence will be useful for understanding the metabolic pathways involved in phenol degradation.

Evaluation of Adjunctive Use of Low-Level Diode Laser Biostimulation with Combined Orthodontic Regenerative Therapy.

BACKGROUND: The combined orthodontic regenerative therapy approach can greatly enhance periodontal conditions and dentofacial aesthetics in many situations. The purpose of this study was to evaluate the effectiveness of 940nm low-level diode laser biostimulation in enhancement of intrabony wound healing with combined orthodontic/regenerative therapy in chronic periodontitis subjects suffering from malocclusion. METHODS: Fifteen chronic periodontitis adult patients with at least two intrabony defects and requiring orthodontic treatment for abnormalities in occlusion were included. A total of 30 defects were divided into two groups and treated in a split mouth design. The defects were treated with combined orthodontic regenerative therapy with laser irradiation (Group I: test group) or with combined orthodontic regenerative therapy alone (Group II: control group). The following hard and soft tissue measurements were recorded at baseline (prior to surgery) and after six and nine months postoperatively: probing depth (PD), clinical attachment level (CAL) by periodontal calibrated probe and bone density (BD) using the DBS-Win software. RESULTS: Probing depth reduction was 64.57%±9.37 and 64.95%±10.07 with no statistically significant difference between Group I and Group II. Percent change in clinical attachment level gain were 59.77%±12.107and 38.83%±7.56 in Group I and II respectively, with a statistically signifi cant difference (P-value =0.005considered signifi cant). Moreover, defects treated with combined orthodontic regenerative therapy with laser irradiation showed signifi cant preservation of bone density with a percent decrease of 4.14%±3.17 at the end of the study period. CONCLUSION: Improvements in clinical and radiographic parameters were observed following the adjunctive use of low-level diode laser therapy and orthodontic regenerative therapy for the management of intrabony defects in chronic periodontitis patients.

Spidroin in carbopol-based gel promotes wound healing in earthworm's skin model.

Spider silk's regenerative, biocompatible, and antimicrobial properties render it a promising biomaterial for wound healing promotion. Spidroin as the main protein component of spider silks was used in this study to evaluate the potential effects on wound healing via topical application of novel spidroin-containing carbopol 934 (CP934) gel. Spidroin was extracted, formulated into CP934 gel, and characterized both in vitro and in vivo. Spidroin gel was translucent and brownish-yellow in color. An optimum viscosity was obtained at 0.6% CP934 at neutral pH. Optimized spidroin gel (0.6% CP934) effectively inhibited the growth of clinical bacterial isolates of methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA) and Escherichia coli at 440 µg/mL with MIC values of 0.98, 4.6, and 8.2 µg/mL, respectively. Optimized spidroin gel was evaluated for wound healing via topical application on wounds surgically induced in Allolobophora caliginosa earthworms used as a robust human skin model. After application for three consecutive days, dramatic reductions in wound closure and reepithelialization duration were observed macroscopically and via histological studies (light and electron microscopy) when compared with control. In conclusion, these results show that spidroin gel is a promising promoter for wound healing, and further studies would be directed toward investigating mechanisms underlying this effect.

Toxicological and epidemiological studies of scorpion sting cases and morphological characterization of scorpions (Leiurusquin questriatus and Androctonus crassicauda) in Luxor, Egypt.

BACKGROUND AND AIM: Scorpion envenomation is a common acute life threatening health problem in developing countries as Egypt. Scorpion venom is a complex structure composed of neurotoxic proteins, salts, acidic proteins, and organic compounds, thereby having of neurologic, cardiovascular, hematologic, and renal side effects, in addition to local effects such as redness, pain, burning, and swelling. AIMS: The study consisted of two parts to describe demographic characteristics, toxicological manifestations of scorpion sting cases in Luxor, Egypt and morphological characterization of the commonest scorpion species in the studied community for easy rapid identification of scorpion species by doctors from the patient history to provide better choices of management. MATERIALS & METHODS: First part of the study include all cases of scorpion envenomation attending General Hospital of Esna, Luxor, Egypt during the period of scorpion activity 2017, demographic data was collected then cases were assessed for clinical manifestations, ECG, complications and deaths related to scorpion species. Second part: scorpions were collected from areas around patient housing for morphological description of their sense organs and venom apparatus by Scanning Electron Microscopy. RESULTS: A total of 110 cases of scorpion stings were reviewed, males 81 (72.9%) and females 30 (27.1%), with a mean age of 31.9 ± 17.9 years. Localized pain was the most frequent presenting complaint (89 80.2%), vomiting was the commonest clinical symptom (90.9%). All scorpion stings were due to the most endemic species in North Africa, Leiurusquinquestriatus & Androctonuscrassicauda. Death rate among cases was (5.5%) all were Abroug's Grade III. Cardiac dysrhythmia and subsequent pulmonary edema were the commonest cause of death. Clinical manifestations were more sever in Leiurus quinquestriatus stings while death rate was more in Androctonus crassicauda stings. The morphological characterization of the sense organs (eyes, pedipalps, pectines& sensory setae) and venom apparatus of the scorpions L. quinquestriatus & A. crassicauda were described by Scanning Electron Microscopy. CONCLUSION: Scorpion sting is an acute heath threating in Southern Egypt, Leiurus quinquestriatus & Androctonus crassicauda are most endemic scorpion species in Southern Egypt. More attention for scorpion envenomation in such subtropical hyper desert localities is required to eradicate toxic scorpion species and prevent possible causes of deaths.

Draft Genome Sequence of the Phenol-Degrading Bacterium Cupriavidus sp. Strain P-10, Isolated from Trichloroethene-Contaminated Aquifer Soil.

A batch culture was enriched on phenol with trichloroethene-contaminated aquifer soil as an inoculum. Cupriavidus sp. strain P-10 was isolated from the culture using a diluted plating method. Here, we report the draft genome sequence and annotation of strain P-10, which provides insights into the metabolic processes of phenol degradation.

Schistosoma, other helminth infections, and associated risk factors in preschool-aged children in urban Tanzania.

BACKGROUND: Despite the high prevalence of helminth infections among preschool-aged children, control programs in sub-Saharan countries primarily focus on school-aged populations. We assessed the prevalence of helminth infections and determined risk factors for infection among preschool-aged children in the urban setting of Dar es Salaam, Tanzania. METHODOLOGY: Starting in October 2015, we conducted a 12-month prospective study among tuberculosis (TB)-exposed children under the age of 5 years and unexposed controls from neighboring households. At the time of recruitment, we collected medical histories, assessed development and cognitive functions, and performed medical examinations. We performed full blood cell counts and screened for HIV and malaria. Point-of-care circulating cathodic antigen (POC-CCA), urine filtration, Kato-Katz, FLOTAC, and Baermann tests were employed to detect helminth infections in urine and stool. Helminth infections were stratified for Schistosoma and other helminths to identify risk factors, using logistic regression. PRINCIPAL FINDINGS: We included 310 children with a median age of 26 months (inter quartile range 17-42 months) in the study. Among these, 189 were TB-exposed and 121 TB-unexposed. Two thirds of the children were anemic (hemoglobin level <11 g/dl) and the HIV prevalence was 1.3%. Schistosoma spp. was the predominant helminth species (15.8%; 95% confidence interval [CI] 12.1-20.3%). Other helminth infections were less frequent (9.0%, 95% CI 6.3-12.8%). Poor hygiene, use of household water sources, and TB-exposure were not associated with helminth infection. Development and cognitive scores did not significantly differ in helminth-infected and uninfected peers, but hemoglobin levels were significantly lower in helminth-infected children (10.1 g/dl vs. 10.4 g/dl, p = 0.027). CONCLUSIONS/SIGNIFICANCE: In Dar es Salaam, a city with more than 4 million inhabitants, the prevalence of Schistosoma spp. infection among preschool-aged children was unexpectedly high. Setting-specific interventions that target preschool-aged children and urban settlements should be considered to reduce the transmission of Schistosoma and other helminth infections and to improve children's health.

Draft Genome Sequence of Pseudomonas sp. LAB-08 Isolated from Trichloroethene-Contaminated Aquifer Soil.

Pseudomonas sp. LAB-08 was isolated from a phenol-fed bioreactor constructed with contaminated aquifer soil as the inoculum. Strain LAB-08 utilized phenol as a sole carbon and energy source. Here, we report the genome sequence and annotation of Pseudomonas sp. LAB-08.

Interspecies interactions are an integral determinant of microbial community dynamics.

This study investigated the factors that determine the dynamics of bacterial communities in a complex system using multidisciplinary methods. Since natural and engineered microbial ecosystems are too complex to study, six types of synthetic microbial ecosystems (SMEs) were constructed under chemostat conditions with phenol as the sole carbon and energy source. Two to four phenol-degrading, phylogenetically and physiologically different bacterial strains were used in each SME. Phylogeny was based on the nucleotide sequence of 16S rRNA genes, while physiologic traits were based on kinetic and growth parameters on phenol. Two indices, J parameter and "interspecies interaction," were compared to predict which strain would become dominant in an SME. The J parameter was calculated from kinetic and growth parameters. On the other hand, "interspecies interaction," a new index proposed in this study, was evaluated by measuring the specific growth activity, which was determined on the basis of relative growth of a strain with or without the supernatant prepared from other bacterial cultures. Population densities of strains used in SMEs were enumerated by real-time quantitative PCR (qPCR) targeting the gene encoding the large subunit of phenol hydroxylase and were compared to predictions made from J parameter and interspecies interaction calculations. In 4 of 6 SEMs tested the final dominant strain shown by real-time qPCR analyses coincided with the strain predicted by both the J parameter and the interspecies interaction. However, in SMEII-2 and SMEII-3 the final dominant Variovorax strains coincided with prediction of the interspecies interaction but not the J parameter. These results demonstrate that the effects of interspecies interactions within microbial communities contribute to determining the dynamics of the microbial ecosystem.

Intraoperative ablation for small HCC not amenable for percutaneous radiofrequency ablation in Child A cirrhotic patients.

BACKGROUND: Radiofrequency ablation (RFA) was initially started by radiologists as a percutaneous treatment, but surgeons started to use RFA by surgical approach for patients with tumors at locations difficult for the percutaneous procedure. The aim was to evaluate the results of intraoperative RFA for small hepatocellular carcinomas (HCCs) (<3 cm) in locations difficult for a percutaneous approach. METHODS: Two hundred forty-seven patients with small solitary HCC (<3 cm) were treated; 196 via percutaneous RFA while 51 patients presented at sites not amenable for percutaneous route. Twenty-seven out of 51 patients underwent surgical resection, while 24/51 patients underwent intraoperative RFA. RESULTS: The location and depth of the tumor from the liver capsule was the only significant factors in the choice of the surgeon between resection and RFA. RFA was successful in all tumors (complete ablation rate of 100 %). In the surgery group, all patients achieved R0 resection. Complication rate was comparable (p = 1.0). After a median follow-up of 37 months (range, 10-45 months), no tumors showed neither local progression nor local recurrence and no significant difference was observed between two groups as regards early recurrence and number of de novo lesions (p = 0.49). One-year and 3-year survival rates were 93 % and 81 %, respectively, in the resection group comparable to the corresponding rates of 92 % and 74 % in the RFA group (p = 0.9). CONCLUSION: For small HCC in locations difficult for a percutaneous approach, intraoperative RFA can be an alternative option for deep-seated tumors necessitating more than one segmentectomy achieving similar tumor control, and overall and disease-free survival.

Nondaily, light daily, and moderate-to-heavy cigarette smokers in a rural area of Egypt: a population-based survey.

INTRODUCTION: Understanding tobacco use among nondaily and light daily cigarette smokers is needed because they are dissimilar from other smokers and may require specific prevention efforts. METHOD: We compared three groups of adult male smokers in rural Egypt: light daily and nondaily smokers versus moderate-to-heavy daily smokers. Data were obtained from a household survey in 2003 in six randomly selected villages in the Nile Delta region of Egypt. There were 7,657 adult participants, of whom 48.6% were male and 51.4% were female. Among them, 1,401 males (37.6%) and 5 females (0.1%) were self-identified as current cigarette smokers; further analysis focused on males. We restricted data analysis to those who reported a stable pattern of 3+ years of smoking. There were 42 nondaily smokers. Daily smokers were subdivided into two groups: light daily smokers (who smoked no more than 10 cigarettes/day; n = 223) and moderate-to-heavy daily smokers (who smoked at least 11 cigarettes/day; n = 769). RESULTS: We found statistically significant differences between these groups on nearly every measure: nondaily smokers tended to be younger and unmarried, but they also had higher levels of education and professional occupations compared with the other smokers. Nondaily and the light daily smokers were more likely than moderate-to-heavy smokers to be planning to quit and to have self-efficacy for quitting, and they were less likely to be smoking in the presence of their wife and children at home. DISCUSSION: Further understanding of nondaily and light daily smokers may aid in tailoring specific interventions.

Transmission of hepatitis C virus between parents and children.

Egyptian children with infected parents are at high risk of infection with hepatitis C (HCV). Analysis of data collected during surveys of rural communities show children whose parents had antibodies to HCV (anti-HCV) were at higher risk for having anti-HCV than children whose parents did not. The association was greater with mothers than fathers and when the parent had HCV RNA. For instance, 87 (14%) of 612 children had anti-HCV whose mothers had HCV RNA compared with 28 (7%) of 401 whose mothers only had anti-HCV and 79 (2.6%) of 3,086 whose mothers were seronegative. These associations persisted after controlling for age, parenteral exposures, and serologic status of the other parent. Sequencing isolates from 13 families with parent(s) and children having HCV RNA showed 10 of 18 had genetically similar viruses. These findings suggest Egyptian children are at high risk of being infected with HCV by their parents and identification of the transmission routes would allow for preventive measures.